Stimuli‐responsive materials have received considerable attention for their application in biomedical fields. Herein, the temperature‐dependent phase transition behavior of acetylated polyrotaxanes (Ac‐PRXs) consisting of acetylated cyclodextrins threaded onto a linear axle polymer in aqueous solution is investigated. The aqueous solutions of Ac‐PRXs exhibit temperature‐dependent transmittance changes when the degree of acetylation is 30–40%. Upon increasing the temperature, Ac‐PRXs are dehydrated and acetyl groups are subsequently associated through hydrophobic interactions. Interestingly, the Ac‐PRXs form coacervate droplets with a diameter of ≈2–3 µm above their cloud points. These temperature‐responsivities of Ac‐PRXs are observed for other PRXs with different axle polymer molecular weights and compositions. Consequently, the acetylation of PRXs is an attractive chemical modification for yielding temperature‐responsivities, and Ac‐PRXs can be applied as temperature‐responsive supramolecular materials.